Separation of calcium channel current components in mouse chromaffin cells superfused with low- and high-barium solutions

Abstract

This study was carried out to characterize the set of voltage-dependent Ca2+ channel subtypes expressed by mouse adrenal chromaffin cells superfused with solutions containing low (2 mM) or high (10 mM) Ba2+ concentrations. Using 50-ms test pulses at 0 mV from a holding potential of -80 mV, averaged peak current in 10 mM Ba2+ was around 1 nA, and in 2 mM Ba2+ 0.36 nA. When using 2 mM Ba2+ as the charge carrier, nifedipine (3 microM) blocked IBa by 40-45%. omega-Conotoxin GVIA (1 microM) caused 26% inhibition, while omega-conotoxin MVIIC (3 microM) produced a 48% blockade. At low concentrations (20 nM), omega-agatoxin IVA caused 5-15% of current inhibition, while 2 microM gave rise to a 35-40% blockade. In 10 mM Ba2+, the blocking effects of nifedipine (40%) and omega-conotoxin GVIA (25%) were similar to those seen in 2 mM Ba2+. In contrast, blockade by omega-conotoxin MVIIC was markedly reduced in 10 mM Ba2+ (20-25%) as compared to 10 mM Ba2+ (48%). The blocking actions of omega-agatoxin IVA (2 microM) were also slowed down in 10 mM Ba2+, though the final blockade was unaffected. In 2 mM Ba2+, IBa was quickly inhibited by over 94% with combined nifedipine + omega-conotoxin MVIIC + omega-conotoxin GVIA; in 10 mM Ba2+, IBa was blocked by 70% with this combination. The data suggest that mouse chromaffin cells express L-type (40%) as well as non-L-type (60%) high-threshold voltage-dependent Ca2+ channels. The current carried by non-L-type Ca2+ channels consists of about 25% N-type and 35% P/Q-type; P-type channels, if anything, are poorly expressed. The data also indicate that the fraction of current blocked by omega-conotoxin MVIIC and omega-agatoxin IVA might considerably change as a function of the Ba2+ concentration of the extracellular solution; taking this fact into consideration, it seems that a residual R-type current is not expressed in mouse chromaffin cells.